Green fluorescent protein as an efficient selection marker for Agrobacterium rhizogenes mediated carrot transformation |
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Authors: | R Baranski E Klocke G Schumann |
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Institution: | (1) Department of Genetics, Plant Breeding and Seed Science, Faculty of Horticulture, Agricultural University of Krakow, Al. 29 Listopada 54, 31-425 Krakow, Poland;(2) Institute of Horticultural Crops, Federal Centre for Breeding Research on Cultivated Plants, Neuer Weg 22/23, 06484 Quedlinburg, Germany |
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Abstract: | Agrobacterium rhizogenes mediated transformation combined with a visual selection for green fluorescent protein (GFP) has been applied effectively
in carrot (Daucus carota L.) transformation. Carrot root discs were inoculated with A4, A4T, LBA1334 and LBA9402 strains, all bearing gfp gene in pBIN-m-gfp5-ER. The results indicate that transformed adventitious roots can be visually selected solely based on
GFP fluorescence with a very high accuracy. The method requires no selection agents like antibiotics or herbicides and enables
a reduction of labour and time necessary for tissue culture. Moreover, individual transformants can be easily excised from
the host tissue and cultured separately. All of the 12 used carrot cultivars produced transformed adventitious roots and the
frequency of discs producing GFP expressing adventitious roots varied from 13 to 85%. The highest transformation rate was
found for A4T and LBA1334 strains possessing chromosomal background of A. tumefaciens C58. The results encourage that visual selection of transformed, fluorescing adventitious roots can be highly effective and
applied routinely for the production of carrot transgenic plants. |
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Keywords: | Agrobacterium rhizogenes Daucus carota GFP Transformation efficiency |
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